Person: Campuzano Ruiz, Susana
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First Name
Susana
Last Name
Campuzano Ruiz
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Química Analítica
Area
Química Analítica
Identifiers
18 results
Search Results
Now showing 1 - 10 of 18
Item Amperometric Immunosensing Scaffolds for Rapid, Simple, Non-Invasive and Accurate Determination of Protein Biomarkers of Well-Accepted and Emerging Clinical Importance(Proceedings, 2017) Pedrero Muñoz, María; Muñoz San Martín, Cristina; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Vargas Orgaz, Eva; Manuel de Villena Rueda, Francisco Javier; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana; Pingarrón Carrazón, José ManuelItem Toward Liquid Biopsy: Determination of the Humoral Immune Response in Cancer Patients Using HaloTag Fusion Protein-Modified Electrochemical Bioplatforms(Analytical Chemistry, 2016) Garranzo Asensio, María; Guzmán Aránguez, Ana Isabel; Povés Francés, Carmen; Fernández Aceñero, Mª Jesús; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Domínguez Muñóz, Gemma; San Frutos Llorente, Luis; Rodríguez Salas, Nuria; Villalba Díaz, Mayte; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Barderas Manchado, RodrigoAutoantibodies raised against tumor-associated antigens have shown high promise as clinical biomarkers for reliable diagnosis, prognosis, and therapy monitoring of cancer. An electrochemical disposable biosensor for the specific and sensitive determination of p53-specific autoantibodies has been developed for the first time in this work. This biosensor involves the use of magnetic microcarriers (MBs) modified with covalently immobilized HaloTag fusion p53 protein as solid supports for the selective capture of specific autoantibodies. After magnetic capture of the modified MBs onto screen-printed carbon working electrodes, the amperometric signal using the system hydroquinone/H2O2 was related to the levels of p53-autoantibodies in the sample. The biosensor was applied for the analysis of sera from 24 patients with high-risk of developing colorectal cancer and 6 from patients already diagnosed with colorectal (4) and ovarian (2) cancer. The developed biosensor was able to determine p53 autoantibodies with a sensitivity higher than that of a commercial standard ELISA using a just-in-time produced protein in a simpler protocol with less sample volume and easily miniaturized and cost-effective instrumentation.Item Multiplexed biosensing diagnostic platforms detecting autoantibodies to tumor-associated antigens from exosomes released by CRC cells and tissue samples showed high diagnostic ability for colorectal cancer(Engineering, 2021) Montero Calle, Ana; Aranguren Abeigon, Itziar; Garranzo Asensio, María; Povés Francés, Carmen; Fernández Aceñero, María Jesús; Martínez Useros, Javier; Sanz, Rodrigo; Dziaková, Jana; Rodríguez Cobos, Javier; Solís Fernández, Guillermo; Povedano, Eloy; Gamella Carballo, Maria; Torrente Rodríguez, Rebeca Magnolia; Alonso Navarro, Miren; Ríos, Vivian de los; Casal, J. Ignacio; Domínguez Muñóz, Gemma; Guzmán Aránguez, Ana Isabel; Peláez García, Alberto, Alberto; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Barderas Manchado, RodrigoColorectal cancer (CRC) is the second leading cause of cancer-related death worldwide. The 5-year survival rate of CRC patients depends on the stage at diagnosis, being higher than 80% when CRC is diagnosed in the early stages but lower than 10% when CRC is diagnosed in advanced stages. Autoantibodies against specific CRC autoantigens (tumor-associated antigens (TAAs)) in the sera of patients have been widely demonstrated to aid in early diagnosis. Thus, we herein aim to identify autoantigens target of autoantibodies specific to CRC that possess a significant ability to discriminate between CRC patients and healthy individuals by means of liquid biopsy. To that end, we examined the protein content of the exosomes released by five CRC cell lines and tissue samples from CRC patients by means of immunoprecipitation coupled with mass spectrometry analysis. A total of 103 proteins were identified as potential autoantigens specific to CRC. After bioinformatics and meta-analysis, we selected 15 proteins that are more likely to be actual CRC autoantigens in order to evaluate their role in CRC prognosis by Western blot (WB) and immunohistochemistry (IHC). We found dysregulation at the protein level for 11 of these proteins in both tissue and plasma exosome samples from patients, along with an association of nine of these proteins with CRC prognosis. After validation, all but one showed a statistically significant high diagnostic ability to distinguish CRC patients and individuals with premalignant lesions from healthy individuals, either by luminescence Halotag-based beads, or by a multiplexed biosensing platform involving the use of magnetic microcarriers as solid support modified with covalently immobilized Halotag fusion proteins constructed for CRC detection. Taken together, our results highlight the usefulness of the approach defined here to identify the TAAs specific to chronic diseases; they also demonstrate that the measurement of autoantibody levels in plasma against the TAAs identified here could be integrated into a point-of-care (POC) device for CRC detection with high diagnostic ability.Item p53 and p63 Proteoforms Derived from Alternative Splicing Possess Differential Seroreactivity in Colorectal Cancer with Distinct Diagnostic Ability from the Canonical Proteins(Cancers, 2023) Montero Calle, Ana; Garranzo Asensio, María; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Poves, Carmen; Dziakova, Jana; Sanz, Rodrigo; Díaz Del Arco, Cristina; Pingarrón Carrazón, José Manuel; Fernández Aceñero, María Jesús; Campuzano Ruiz, Susana; Barderas Manchado, RodrigoColorectal cancer (CRC) is the third most common cancer and the second most frequent cause of cancer-related death worldwide. The detection in plasma samples of autoantibodies against specific tumor-associated antigens has been demonstrated to be useful for the early diagnosis of CRC by liquid biopsy. However, new studies related to the humoral immune response in cancer are needed to enable blood-based diagnosis of the disease. Here, our aim was to characterize the humoral immune response associated with the different p53 and p63 proteoforms derived from alternative splicing and previously described as aberrantly expressed in CRC. Thus, here we investigated the diagnostic ability of the twelve p53 proteoforms and the eight p63 proteoforms described to date, and their specific N-terminal and C-terminal end peptides, by means of luminescence HaloTag beads immunoassays. Full-length proteoforms or specific peptides were cloned as HaloTag fusion proteins and their seroreactivity analyzed using plasma from CRC patients at stages I-IV (n = 31), individuals with premalignant lesions (n = 31), and healthy individuals (n = 48). p53γ, Δ40p53β, Δ40p53γ, Δ133p53γ, Δ160p53γ, TAp63α, TAp63δ, ΔNp63α, and ΔNp63δ, together with the specific C-terminal end α and δ p63 peptides, were found to be more seroreactive against plasma from CRC patients and/or individuals with premalignant lesions than from healthy individuals. In addition, ROC (receiver operating characteristic) curves revealed a high diagnostic ability of those p53 and p63 proteoforms to detect CRC and premalignant individuals (AUC higher than 85%). Finally, electrochemical biosensing platforms were employed in POC-like devices to investigate their usefulness for CRC detection using selected p53 and p63 proteoforms. Our results demonstrate not only the potential of these biosensors for the simultaneous analysis of proteoforms’ seroreactivity, but also their convenience and versatility for the clinical detection of CRC by liquid biopsy. In conclusion, we here show that p53 and p63 proteoforms possess differential seroreactivity in CRC patients in comparison to controls, distinctive from canonical proteins, which should improve the diagnostic panels for obtaining a blood-based biomarker signature for CRC detection.Item Multiplexed monitoring of a novel autoantibody diagnostic signature of colorectal cancer using HaloTag technology-based electrochemical immunosensing platform(Theranostics, 2020) Garranzo Asensio, María; Guzmán Aránguez, Ana Isabel; Povedano, Eloy; Ruiz Valdepeñas Montiel, Víctor; Povés Francés, Carmen; Fernández Aceñero, María Jesús; Montero Calle, Ana; Solís Fernández, Guillermo; Fernández Díez, Servando; Camps, Jordi; Arenas, Meritxell; Rodríguez Tomás, Elisabeth; Joven, Jorge; Sánchez Martínez, Maricruz; Rodrígez, Nuria; Domínguez Muñóz, Gemma; Yáñez Sedeño, Paloma; Pingarrón Carrazón, José Manuel; Campuzano Ruiz, Susana; Barderas Manchado, RodrigoBackground and Purpose: The humoral immune response in cancer patients can be used for early detection of the disease. Autoantibodies raised against tumor-associated antigens (TAAs) are promising clinical biomarkers for reliable cancer diagnosis, prognosis, and therapy monitoring. In this study, an electrochemical disposable multiplexed immunosensing platform able to integrate difficult- and easy-to-express colorectal cancer (CRC) TAAs is reported for the sensitive determination of eight CRC-specific autoantibodies. Methods: The electrochemical immunosensing approach involves the use of magnetic microcarriers (MBs) as solid supports modified with covalently immobilized HaloTag fusion proteins for the selective capture of specific autoantibodies. After magnetic capture of the modified MBs onto screen-printed carbon working electrodes, the amperometric responses measured using the hydroquinone (HQ)/H2O2 system were related to the levels of autoantibodies in plasma. Results: The biosensing platform was applied to the analysis of autoantibodies against 8 TAAs described for the first time in this work in plasma samples from healthy asymptomatic individuals (n=3), and patients with high-risk of developing CRC (n=3), and from patients already diagnosed with colorectal (n=3), lung (n=2) or breast (n=2) cancer. The developed bioplatform demonstrated an improved discrimination between CRC patients and controls (asymptomatic healthy individuals and breast and lung cancer patients) compared to an ELISA-like luminescence test. Conclusions: The proposed methodology uses a just-in-time produced protein in a simpler protocol, with low sample volume, and involves cost-effective instrumentation, which could be used in a high-throughput manner for reliable population screening to facilitate the detection of early CRC patients at affordable cost.Item Fast and sensitive diagnosis of autoimmune disorders through amperometric biosensing of serum anti-dsDNA autoantibodies(Biosensors & Bioelectronics, 2020) López Ruíz, Beatriz ; Arévalo Pérez, Beatriz; Serafín González-Carrato, Verónica; Sánchez-Paniagua López, Marta; Montero Calle, Ana; Barderas Manchado, Rodrigo; López Ruiz, María Beatriz; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José ManuelThis work reports the first amperometric biosensor involving the use of neutravidin-functionalized magnetic microbeads (NA-MBs) modified with a biotinylated-anti-dsDNA (b-dsDNA) as efficient magnetic microcarriers to selectively capture anti-dsDNA autoantibodies (IgG, IgA and IgM AAbs) present in the sera of patients with rheumatoid arthritis (RA). Subsequently, the attached anti-dsDNA AAbs are detected with a mixture of conventional HRP-labeled secondary antibodies (HRP-anti-human IgG/IgM/IgA mixture). The biorecognition event is monitored by amperometric transduction using the hydroquinone (HQ)/H2O2 system upon capturing the modified MBs on the surface of screen-printed carbon electrodes (SPCEs). The developed bioplatform exhibits a linear calibration plot ranging from 1 to 200 IU mL−1 with a LOD of 0.3 IU mL−1 for anti-dsDNA AAbs standards. In addition, the biosensor allows performing the determination of the anti-dsDNA AAbs levels directly in 100-times diluted serum samples from patients diagnosed with RA and in just 75 min. The obtained results are in agreement with those provided by an ELISA kit and allow discrimination between positive and negative samples.Item Disposable Amperometric Immunosensor for the Determination of Human P53 Protein in Cell Lysates Using Magnetic Micro-Carriers(Biosensors, 2016) Pedrero Muñoz, María; Manuel de Villena Rueda, Francisco Javier; Muñoz San Martín, Cristina; Campuzano Ruiz, Susana; Garranzo Asensio, María; Barderas Manchado, Rodrigo; Pingarrón Carrazón, José ManuelAn amperometric magnetoimmunosensor for the determination of human p53 protein is described in this work using a sandwich configuration involving the covalent immobilization of a specific capture antibody onto activated carboxylic-modified magnetic beads (HOOC-MBs) and incubation of the modified MBs with a mixture of the target protein and horseradish peroxidase-labeled antibody (HRP-anti-p53). The resulting modified MBs are captured by a magnet placed under the surface of a disposable carbon screen-printed electrode (SPCE) and the amperometric responses are measured at −0.20 V (vs. an Ag pseudo-reference electrode), upon addition of hydroquinone (HQ) as a redox mediator and H2O2 as the enzyme substrate. The magnetoimmunosensing platform was successfully applied for the detection of p53 protein in different cell lysates without any matrix effect after a simple sample dilution. The results correlated accurately with those provided by a commercial ELISA kit, thus confirming the immunosensor as an attractive alternative for rapid and simple determination of this protein using portable and affordable instrumentation.Item Disposable electrochemical immunoplatform to shed light on the role of the multifunctional glycoprotein TIM-1 in cancer cells invasion(Talanta, 2024) Quinchia, Jennifer; Blázquez-García, Marina; Torrente Rodríguez, Rebeca Magnolia; Ruiz Valdepeñas Montiel, Víctor; Serafín González-Carrato, Verónica; Rejas-González, Raquel; Montero-Calle, Ana; Orozco, Jahir; Pingarrón Carrazón, José Manuel; Barderas Manchado, Rodrigo; Campuzano Ruiz, SusanaDetecting overexpression of cancer biomarkers is an excellent tool for diagnostic/prognostic and follow-up of patients with cancer or their response to treatment. This work illustrates the relevance of interrogating the levels of T-cell immunoglobulin and mucin domain 1 (TIM-1) protein as a diagnostic/prognostic biomarker of high-prevalence breast and lung cancers by using an amperometric disposable magnetic microparticles-assisted immunoplatform. The developed method integrates the inherent advantages of carboxylic acid-functionalized magnetic beads (HOOC-MBs) as pre-concentrator support and the amperometric transduction at screen-printed carbon electrodes (SPCEs). The immunoplatform involves a sandwich-type immunoassay assembled on HOOC-MBs through the specific capture/labeling of TIM-1 using capture antibodies and horseradish peroxidase (HRP)-conjugated biotinylated detection antibodies as biorecognition elements. The magnetic immunoconjugates were confined onto the working electrode (WE) surface of the SPCEs for amperometric detection using the hydroquinone/hydrogen peroxide/HRP (HQ/H2O2/HRP) redox system. The method allows the selective detection of TIM-1 protein over the 87–7500 pg mL−1 concentration range in only 45 min, with a limit of detection of 26 pg mL−1. The developed bioplatform was successfully applied to the analysis of breast and lung cancer cell extracts, providing the first quantitative results of the target glycoprotein in these types of samples.Item Click chemistry-assisted antibodies immobilization for immunosensing of CXCL7 chemokine in serum(Journal of Electroanalytical Chemistry, 2019) Guerrero Irigoyen, Sara; Agüí Chicharro, María Lourdes; Barderas Manchado, Rodrigo; Campuzano Ruiz, Susana; Yáñez-Sedeño Orive, Paloma; Pingarrón Carrazón, José Manuel; Cadanno Mendía, DonaThe first electrochemical immunosensor for the determination of CXCL7 (chemokine (C-X-C motif) ligand 7) autoimmune biomarker is reported in this work. Click chemistry-assisted antibodies immobilization was per formed by reaction of azide functionalized-multi-walled carbon nanotubes (MWCNTs) and ethynyl-IgG onto screen-printed carbon electrodes. The capture antibodies were further immobilized onto IgG-MWCNTs con jugates. After a blocking step with casein, a sandwich immunoassay was implemented involving biotinylated detector antibodies and alkaline phosphatase (AP)-streptavidin conjugate. Differential pulse voltammetry upon addition of 1-naphthylphosphate was used as the analytical readout. A linear calibration plot between 0.5 and 600 pg mL−1 CXCL7 and a LOD value of 0.1 pg mL−1 were obtained. The usefulness of the immunosensor was demonstrated by the successful analysis of serum samples from patients with rheumatoid arthritis.Item Pursuing precision in medicine and nutrition: the rise of electrochemical biosensing at the molecular level(Analytical and Bioanalytical Chemistry, 2024) Campuzano Ruiz, Susana; Barderas Manchado, Rodrigo; Moreno-Casbas, María Teresa; Almeida, Ángeles; Pingarrón Carrazón, José ManuelIn the era that we seek personalization in material things, it is becoming increasingly clear that the individualized management of medicine and nutrition plays a key role in life expectancy and quality of life, allowing participation to some extent in our welfare and the use of societal resources in a rationale and equitable way. The implementation of precision medicine and nutrition are highly complex challenges which depend on the development of new technologies able to meet important requirements in terms of cost, simplicity, and versatility, and to determine both individually and simultaneously, almost in real time and with the required sensitivity and reliability, molecular markers of different omics levels in biofluids extracted, secreted (either naturally or stimulated), or circulating in the body. Relying on representative and pioneering examples, this review article critically discusses recent advances driving the position of electrochemical bioplatforms as one of the winning horses for the implementation of suitable tools for advanced diagnostics, therapy, and precision nutrition. In addition to a critical overview of the state of the art, including groundbreaking applications and challenges ahead, the article concludes with a personal vision of the imminent roadmap.